Human interleukin-1.beta. (hIL-1.beta.) is produced by stimulated monocytes initially as a 269 amino acid protein with Mr of 30,747. It is proteolytically converted to a biologically active form consisting of the 153 amino acid carboxy-terminal portion from position 117 to position 269, with Mr of 17,377. The 269 amino acid protein is referred to as pro hIL-1.beta.; the 153 amino acid protein is referred to as mature hIL-1.beta..
European Patent Application Publication Number 0165654, filed by Immunex Corporation in March 1985, discloses expression of recombinant mature hIL-1.beta. having a methionine residue preceding position 1 (met hIL-1.beta.). Although the gene encoding hIL-1.beta. was amplified in E. coli, it was expressed in yeast. In June 1985 researchers at Immunex published a paper (March et al., Nature 315:641-647, June 20, 1985) describing expression in E. coli of recombinant mature met hIL-1.beta. using a vector containing the .lambda. phage P.sub.L promoter. This paper states at pages 644-5 that the cells produced high levels of IL-1 activity. There is no disclosure of purification of the protein. In 1986 researchers at Immunex published another paper (Kronheim et al., Biotechnology 4:1078-1082, December 1986) indicating at page 1080 that the "preliminary constructions" described in the March et al. paper did not produce high levels of recombinant hIL-1.beta., and that purified hIL-1.beta. had been obtained previously only from human monocytes. The Kronheim et al. paper describes high-level expression of hIL-1.beta. in E. coli, using expression system pLNIL1.beta., also containing the .lambda. phage P.sub.L promoter, and purification of the protein.
European Patent Application Publication Number 0161901 and Auron et al. Proc. Natl. Acad. Sci. USA 81:7907-7911 (1984) disclose cloning vehicles containing the nucleotide coding sequence for pro hIL-1.beta. (pcD415 and pcD1218) and various fragments thereof. EPA Publication Number 0161901 states that the sequences can be fused into an expression vector and expressed in either eukaryotic or prokaryotic cells, but does not disclose a specific expression system. Dinarello et al. J. Clin. Invest. 77:1734-1739 (June 1986) discloses expression in E. coli of a protein consisting of amino acids 71-269 of pro hIL-1.beta. preceded by 24 amino acids contributed by the expression vector, followed by enzymatic digestion to produce a protein consisting of amino acids 112-269 of pro hIL-1.beta.. The paper states that the expression vector was constructed by inserting a fragment of pcD1218 into an E. coli expression plasmid, which is not further identified. Rossenwasser et al., Proc. Natl. Acad. Sci. USA, 83:5243-5246 (July 1987) discloses expression in COS monkey cells of pro hIL-1.beta. using plasmids pcD415 and 1218, and various deleted forms, including one consisting of about 136 amino acids of the carboxy terminal of mature hIL-1.beta..
Pro human interleukin 1.alpha. (pro hIL-1.alpha.) is a 271 amino acid protein with about 27% homology to pro hIL-1.beta.. It is processed proteolytically to a biologically active 17 approximately kD protein consisting of the 159 amino acids of the carboxy-terminal of pro hIL-1.alpha.. Gubler et al., J. Immunology 136:2492-2497 (1986) discloses expression in E. coli of the carboxy-terminal 154 amino acids of pro hIL-1.alpha., using a plasmid containing the .lambda. phage P.sub.L promoter.
Plasmid vectors containing lac promoters such as pUC8 have been described (Vieira and Messing et al., Gene 19:259-268 (1982)) and are available commercially from Pharmacia, Piscataway, NJ and Bethesda Research Laboratory, Bethesda MD. EPA Publication 0161901 and the Rossenwasser et al. publication cited above mention pUC8 in connection with construction of cloning and expression vectors, but only as a source of a polylinker fragment used in constructing plasmid pcD1218.